Crystallization and preliminary X-ray diffraction study of the bacterially expressed Fv from the monoclonal anti-lysozyme antibody D1.3 and of its complex with the antigen, lysozyme

Two mouse monoclonal anti-anti-idiotopic antibodies (anti-anti-Id, Ab3), AF14 and AF52, were prepared by immunizing BALB͞c mice with rabbit polyclonal antiidiotypic antibodies (anti-Id, Ab2) raised against antibody D1.3 (Ab1) specific for the antigen hen egg lysozyme. AF14 and AF52 react with an ''internal image'' monoclonal mouse anti-Id antibody E5.2 (Ab2), previously raised against D1.3, with affinity constants (1.0 ؋ 10 9 M ؊1 and 2.4 ؋ 10 7 M ؊1 , respectively) usually observed in secondary responses against protein antigens. They also react with the antigen but with lower affinity (1.8 ؋ 10 6 M ؊1 and 3.8 ؋ 10 6 M ؊1 ). This pattern of affinities for the anti-Id and for the antigen also was displayed by the sera of the immunized mice. The amino acid sequences of AF14 and AF52 are very close to that of D1.3. In particular, the amino acid side chains that contribute to contacts with both antigen and anti-Id are largely conserved in AF14 and AF52 compared with D1.3. Therapeutic immunizations against different pathogenic antigens using anti-Id antibodies have been proposed. Our experiments show that a response to an anti-Id immunogen elicits anti-anti-Id antibodies that are optimized for binding the anti-Id antibodies rather than the antigen.The complementarity determining regions (CDR) of Ab display great structural diversity and constitute a vast array of potential antigenic determinants. Under appropriate experimental conditions, these determinants give rise to immune responses and are called idiotypic (1, 2). Idiotypes of Ab are the sum of idiotopes (Id) or antigenic determinants characterized experimentally by the reaction of an anti-idiotopic (anti-Id) antibody (Ab2) with the Id (Ab1). In most cases, idiotypes have been shown to be associated, partially or entirely, with the CDR of Ab molecules (3).The observation that external antigens and anti-Id Ab can competitively bind to specific Ab has led to proposals that anti-Id Ab may carry an ''internal image'' (2) of the external antigen. Functional mimicry of ligands of biological receptors by anti-Id Ab has been described in several systems (4). The potential for mimicking external antigens has led to proposals (5, 6) to use anti-idiotypic Ab as surrogate antigens. Recent reviews (7-9) have discussed this topic from a structural viewpoint. Indeed, anti-Id Ab have been used in exploring therapeutic immunizations against different pathogenic antigens.A report from this laboratory compared the threedimensional structures of the complexes between an Ab1-antigen and an Ab1-anti-Id and provided a concrete example of how an anti-Id Ab can structurally mimic an external antigen (10, 11). In this system, the Ab1 was the mAb D1.3 specific for the antigen, hen egg lysozyme (HEL). The three-dimensional crystal structure of the complex between the Fv fragment of D1.3 with HEL has been determined at the resolution of 1.8 Å (12). Subsequently, the three-dimensional structure of a complex between the Fv from D1.3 and the anti-Id Fv from mAb E5.2 (the Ab2; BALB͞c I...

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“…The amino acid sequences of the V H and V L segments of AF14 and AF52, compared with that of D1.3 (16), are shown in Fig. 4.…”

Section: Resultsmentioning

confidence: 99%

Characterization of anti-anti-idiotypic antibodies that bind antigen and an anti-idiotype

Goldbaum

Velikovsky

Dall’Acqua³

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

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“…The complex of the hen lysozyme and an antibody fragment was studied extensively (Amit et al, 1986;Bhat et al, 1990;Boulot et al, 1990). The antibody D1.3 has also been used as a model for affinity maturation by phage selection technology.…”

Section: Kinetics and Affinity Related To Structural Differences In Lmentioning

confidence: 99%

Kinetic analysis of engineered antibody‐antigen interactions

Malmqvist

1994

J of Molecular Recognition

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Molecular engineering of antibodies has made it possible to produce specific domains of the antibody molecule and combine them with other protein domains to achieve new properties. Using site directed mutagenesis, amino acid residues can be exchanged within the binding site; and, by analysis of crystal structures, the positions of these amino acids can be determined in three dimensions at atomic resolution. In addition, gene libraries and phage selection technology can be used to generate new antibody fragments directly from a gene pool. Both mutagenesis and selection from libraries offer opportunities to identify antibody-derived molecules with altered and useful antigen recognition properties. The detailed analysis of both kinetic and equilibrium binding affinity are therefore essential to understand the activity of the molecules resulting from antibody engineering and to guide the progress of their further design. This paper reviews recently evolving techniques for the binding analysis of antibodies, their functional domains and antibody chimerae.

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“…An economic approach to construction of SMAA complexes would be to employ a single 'humanized' MAb specific for a small universal peptide tag antigen, used to present any protein to which the tag antigen was attached. Furthermore, the possible use of Fab or Fv fragments for construction of SMAA complexes would enable a largescale production of these fragments using bacterial expression systems (Boulot et al, 1990;Skerra et al, 1991), thus avoiding a laborious and expensive production of whole human antibodies in rat myeloma cell lines (Reichmann et al, 1988;Verhoeyen et al, 1988;Tempest et al, 1991).…”

Section: Introductionmentioning

confidence: 99%

Construction of solid matrix-antibody-antigen complexes containing simian immunodeficiency virus p27 using tag-specific monoclonal antibody and tag-linked antigen

Hanke

Szawlowski

Randall

1992

Journal of General Virology

139

107

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We have previously shown that immunization with solid matrix-antigen-antibody (SMAA) complexes induces both vigorous humoral and cell-mediated immune responses and have suggested that this method of vaccination may be developed for use in humans, and potentially as a vaccine against AIDS. Here we demonstrate that a small oligopeptide can act as a tag for the construction of SMAA complexes using a tagspecific monoclonal antibody and tag-linked antigens. We show that a 14-amino acid oligopeptide, present in the phospho (P) and V proteins of simian virus 5 (SV5), retains its antigenicity when attached to the C terminus of three 'foreign' proteins [p27 and gp110 of simian immunodeficiency virus (SIV) and glutathione S-transferase] such that these proteins can be incorporated into SMAA complexes using a monoclonal antibody (MAb) that was originally raised against the native SV5 P and V proteins. Mice were immunized with SMAA complexes containing recombinant p27-TAG and MAbs have been isolated that recognized native SIV p27. The significance of these results in terms of the development of SMAA complexes as human vaccines is discussed.

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Crystallization and preliminary X-ray diffraction study of the bacterially expressed Fv from the monoclonal anti-lysozyme antibody D1.3 and of its complex with the antigen, lysozyme

Cited by 58 publications

References 9 publications

Characterization of anti-anti-idiotypic antibodies that bind antigen and an anti-idiotype

Characterization of anti-anti-idiotypic antibodies that bind antigen and an anti-idiotype

Kinetic analysis of engineered antibody‐antigen interactions

Construction of solid matrix-antibody-antigen complexes containing simian immunodeficiency virus p27 using tag-specific monoclonal antibody and tag-linked antigen

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